LOCUS       sequence-39535-e        8467 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8467)
  AUTHORS   Kobayashi T, Ooms LS, Ikizler M, Chappell JD, Dermody TS
  TITLE     An improved reverse genetics system for mammalian orthoreoviruses.
  JOURNAL   Virology. 2010 Mar 15;398(2):194-200. Epub 2009 Dec 29.
  PUBMED    20042210
REFERENCE   2  (bases 1 to 8467)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Virology.";
            date: "2010-03-15"; volume: "398(2)"; pages: "194-200. Epub 2009 Dec
            29"
COMMENT     Sequence Label: pT7-L3-S3T1L
FEATURES             Location/Qualifiers
     source          1..8467
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      4096..4143
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      5582..5629
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(5697..5715)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5698..5715)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6006..6025)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(6236..6255)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6355..6377
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6415..6433)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6501..6605
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6606..6674,6675..7466)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6824..6843)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7637..8225
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8126..8145
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8379..8396
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 atcgatcccg ggaattctaa tacgactcac tatagctaat cgtcaggatg aagcggattc
       61 caaggaagac aaagggcaaa tccagcggaa agggcaatga ctcaacagaa agatcggacg
      121 atggctcgag ccaattacga gacaagcaaa acaataaggc tggccctgcc actacggagc
      181 ctggcacatc taaccgagag caatacagag cccgaccagg tattgcatct gtgcagaggg
      241 ccactgaaag tgcagaactg cccatgaaga ataatgacga agggacgcca gataagaaag
      301 gaaatactag gggcgactta gttaatgagc atagtgaggc taaagacgag gcggatgaag
      361 cgacgcagaa gcaggcaaaa gacacagaca aaagtaaagc gcaagtcaca tattcagaca
      421 ctggtatcaa taatgctaat gaactgtcaa gatctgggaa tgtggataat gagggtggaa
      481 gtaatcagaa gccgatgtcc accagaatag ctgaagcaac gtctactata gtgtcgaaac
      541 atcctgcgcg tgttgggtta ccacctaccg ctagcagtgg tcatgggtat cagtgccacg
      601 tctgttctgc agtcctgttt agtcctttag acctagatgc ccacgtcgcc tcacatggtt
      661 tgcatggtaa catgacatta acatcgagtg agattcagcg acatataact gagttcatca
      721 gctcatggca aaatcatcct attgttcaag tttcggctga tgtcgaaaat aagaagactg
      781 ctcaactgct tcacgctgac actcctcgac tcgtcacttg ggatgctggt ttgtgtactt
      841 cgttcaaaat cgtcccgatt gtgccagctc aggtgccgca ggatgtactg gcctatacgt
      901 ttttcacctc ttcatatgcc atccaatcac cgtttccaga ggcggcagtg tctaggattg
      961 tggtgcatac gagatgggca tctaatgttg actttgaccg agattcgtct gtcatcatgg
     1021 cgccacctac agaaaacaat atccatttgt ttaagcagtt actaaatact gaaaccctgt
     1081 ctgtaagggg ggctaatccg ctaatgttta gagcgaacgt gttgcatatg ttgctggagt
     1141 tcgtattaga taacttgtat ctgaacagac atacgggatt ctctcaagac cacacaccat
     1201 ttactgaggg tgctaatctg cgctcacttc ctggccccga tgctgagaaa tggtactcga
     1261 ttatgtatcc aacgcgcatg ggaacgccga atgtatcaaa aatatgtaat ttcgtcgcct
     1321 cttgtgtgcg aaatcgggtt ggacgatttg atcgagcaca gatgatgaac ggagctatgt
     1381 cagagtgggt ggatgtcttc gagacttcag acgcgcttac cgtctccatt cgaggtcgat
     1441 ggatggctag actagctcgc atgaacataa atccaacaga gatcgaatgg gcgttgactg
     1501 aatgtgcaca aggatatgtg actgtcacaa gtccttacgc tcctagcgta aatagattga
     1561 tgccctatcg tatctccaac gctgagcggc aaatatcaca gataatcagg atcatgaaca
     1621 ttggcaataa cgcgacggtg atacaacctg ttctgcaaga tatttcagta ctccttcaac
     1681 gcatatcacc actccaaata gatccaacta ttatttccaa cactatgtca acagtctcgg
     1741 agtctactac tcagaccctc agccccgcgt cctcaatttt gggtaaacta cgaccgagta
     1801 actcagattt ctctagtttt agagtcgcgt tggctggatg gctttataat ggagttgtga
     1861 cgacggtgat tgatgatagt tcatatccaa aagacggtgg cagcgtgacc tcacttgaaa
     1921 atctgtggga tttcttcatc cttgcgcttg ctctaccact gacaactgac ccctgtgcac
     1981 ctgtgaaagc attcatgacc ttagctaaca tgatggttgg tttcgagaca atccctatgg
     2041 ataatcagat ctatactcaa tcgagacgcg cgagtgcttt ctcaacgcct cacacgtggc
     2101 cacgatgctt catgaatatc cagttaattt ctccaatcga tgctcctatc ttgcgacagt
     2161 gggctgaaat tattcataga tactggccta acccttcaca gattcgttat ggtgcaccga
     2221 acgttttcgg ctcggcaaat ttgttcactc cacctgaggt gctgttattg ccaatcgatc
     2281 atcaaccagc taatgtaaca acgccaacgc tggacttcac caatgagttg actaattggc
     2341 gcgctcgtgt ctgtgagctt atgaagaatc ttgttgataa tcaaagatat caacctggat
     2401 ggacacaaag tctagtctcg tcaatgcgcg gaacgctaga caaattgaag ttgattaaat
     2461 cgatgacacc catgtatctg caacagctgg ctccggtaga gttagcagtg atagctccca
     2521 tgttgccttt cccacctttc caggtgccct acgtccgtct cgatcgtgat agagttccaa
     2581 caatggtcgg agtaacacga cagtcacgag atactattac tcagccagcg ctatcgctgt
     2641 cgacgaccaa tactactgtt ggcgtgccac tagctctaga cgcgagggct atcaccgttg
     2701 cgctgttgtc agggaaatat ccgccggatt tagtgacaaa tgtatggtac gctgatgcca
     2761 tttatccaat gtatgcagac actgaggtgt tctctaatct tcagagagac atgattacct
     2821 gcgaggccgt gcagacatta gtgactctgg tggcgcaaat atcagagacc cagtatcctg
     2881 tagataggta tcttgattgg atcccatcac tgagagcatc ggcggcgacg gcggcgacat
     2941 ttgctgagtg ggttaatact tcaatgaaga cggcgtttga tttgtctgac atgctgttag
     3001 agcctctcct aagcggtgat ccgaggatga ctcaactagc gattcagtat cagcaataca
     3061 atggcagaac gtttaatgtc atacctgaaa tgccaggttc agttattgct gactgcgttc
     3121 aactaacagc agaagtcttc aatcacgaat ataacctgtt tgggattgcg cggggtgata
     3181 tcatcattgg ccgtgttcag tcgacacatt tgtggtcacc gctggctcct ccacctgacc
     3241 tggtgtttga tcgtgacact cctggtgttc acatcttcgg acgagattgc cgcatatcgt
     3301 ttggaatgaa tggcgccgcg ccaatgatta gagatgagac tggaatgatg gtgccttttg
     3361 aaggaaattg gatcttccca ctggcgcttt ggcaaatgaa tacacgatat tttaatcaac
     3421 aattcgacgc gtggattaag acaggagagt tgcgaatccg tattgagatg ggcgcgtatc
     3481 catatatgtt gcattactat gatccacgtc agtacgctaa tgcatggaat ttaacatccg
     3541 cctggcttga agaaattaca ccgacgagca tcccatccgt gcctttcatg gtgcccattt
     3601 caagtgatca tgacatttcc tctgccccag ctgtccaata catcatctcg actgaatata
     3661 atgatcggtc tttattctgc actaactcat catctcccca aactatcgct ggaccagaca
     3721 aacacattcc agttgaaaga tataacattc tgaccaaccc cgacgctcca cccacgcaga
     3781 tacaactgcc tgaagttgtt gacttgtata acgtcgtcac acgctatgcg tatgagactc
     3841 cgcctattac cgctgtcgtt atgggtgttc cttgatcctc atcctcccaa caggtgctag
     3901 agcatcgcgc tcgatgctag ttgggccgat tcatcgggtc ggcatggcat ctccacctcc
     3961 tcgcggtccg acctgggcat ccgaaggagg acgtcgtcca ctcggatggc taagggagag
     4021 ctcaaaaaaa aggatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     4081 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     4141 ttgctgaaag gaggaactat atccggatcg agatcctcta gagtcgacct gcaggcatgc
     4201 aagctttaat acgactcact atagctaaag tcacgcctgt tgtcgtcact atggcttcct
     4261 cactcagggc tgcgatctct aagatcaaga gagatgatgt tggtcagcaa gtttgtccaa
     4321 attatgtcat gcttagatca tcggtcacaa cgaaggtggt acgaaacgtt gttgagtatc
     4381 aaatccgtac aggtggattc ttttcgtgcc tagctatgtt gagaccgctc cagtatgcta
     4441 aacgtgagcg tctgcttgga caaaggaatc tggaacgcat atcgactagg gacattcttc
     4501 agacacgcga tttgcactca ttgtgcatgc caactcctga cgcgccaatg tccaatcatc
     4561 aggcagccac catgagagag ctgatctgca gctacttcaa ggtcgaccac actgatgggt
     4621 tgaaatatat acccatggat gagagatatt ctccatcatc gcttgccaga ttgttcacta
     4681 tgggtatggc tggactacac attaccactg agccttccta caaacgtgtg cccatcatgc
     4741 acttggcagc agatttggac tgcatgacgt tagctttacc ctacatgatt acacttgatg
     4801 gtgacacagt ggtacctgtc gccccaacgc tttctgcaga acagcttttg gatgatggac
     4861 ttaagggatt agcatgcatg gatatctcat acggatgtga ggtggacgct agcaaccgat
     4921 cagctggtga tcagagcatg gattcttcac gatgcatcaa tgagttatat tgcgaggaaa
     4981 cggcagaagc tatctgtgta ctcaaaacat gtcttgtgct gaactgtatg caattcaaac
     5041 ttgagatgga tgatttagca cacaacgcta ctgagctgga caagatacag atgatgatac
     5101 cttttagtga acgtgttttc agaatggctt cttcatttgc taccattgat gcccagtgtt
     5161 tcaggttttg tgtgatgatg aaggataaga atttgaagat agacatgcgt gaaacgatga
     5221 gactttggac tcgatcggcg ttggacgatt cagtggttac gtcatctttg agtatttcgc
     5281 tggatcgagg tcgatgggta gcagctgatg ctactgatgc tagattgctg gtgtttccaa
     5341 ttcgcgtgta atgggtgagt gagccgatgt ggtcgccaag acatgtgccg gtgtcttggt
     5401 ggtgggtggc gcctaatcat cgggtcggca tggcatctcc acctcctcgc ggtccgacct
     5461 gggcatccga aggaggacgt cgtccactcg gatggctaag ggagagctca aaaaaaagga
     5521 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     5581 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
     5641 aactatatcc ggatcgagat cctctagagt cgacctgcag gcatgcaagc ttgtattcta
     5701 tagtgtcacc taaatcgtat gtgtatgata cataaggtta tgtattaatt gtagccgcgt
     5761 tctaacgaca atatgtacaa gcctaattgt gtagcatctg gcttactgaa gcagacccta
     5821 tcatctctct cgtaaactgc cgtcagagtc ggtttggttg gacgaacctt ctgagtttct
     5881 ggtaacgccg ttccgcaccc cggaaatggt cagcgaacca atcagcaggg tcatcgctag
     5941 ccagatcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc
     6001 tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat
     6061 gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc
     6121 catctccttg caccattcct tgcggcggcg gtgctcaacg gcctcaacct actactgggc
     6181 tgcttcctaa tgcaggagtc gcataaggga gagcgtcgat atggtgcact ctcagtacaa
     6241 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     6301 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     6361 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     6421 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     6481 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     6541 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     6601 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     6661 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     6721 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     6781 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     6841 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     6901 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     6961 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     7021 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     7081 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     7141 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     7201 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     7261 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     7321 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     7381 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     7441 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     7501 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     7561 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     7621 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     7681 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     7741 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     7801 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     7861 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     7921 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7981 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     8041 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     8101 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     8161 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     8221 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     8281 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     8341 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     8401 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     8461 gggggat
//
